Secondary Antibodies
Anti-biotin (D5A7) Rabbit mAb #5597
Protocols
- 5597:
- ELISA Peptide, ELISA-P, Western Blotting
Specificity / Sensitivity
Anti-Biotin (D5A7) Rabbit mAb recognizes biotin attached to proteins, peptides, oligonucleotides, or solid matrices.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a biotinylated protein.
Western Blotting
Western Blot analysis of extracts from HeLa and NIH/3T3 cells using S6 Ribosomal Protein (54D2) Mouse mAb (Biotinylated) #3944 followed by Anti-biotin (D5A7) Rabbit mAb and anti-rabbit IgG, HRP linked Antibody #7074.
ELISA-Peptide
ELISA using Anti-biotin (D5A7) Rabbit mAb coated plates to capture biotinylated phospho and non-phosphopeptides. Peptide detection was performed using Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 followed by Anti-mouse IgG, HRP-linked Antibody #7076. TMB Substrate #7004 was added for color development.
ELISA-Peptide
ELISA showing absorbance at 450 nm of biotinylated and non-biotinylated peptides detected with Anti-biotin (D5A7) Rabbit mAb at different concentrations. TMB substrate #7004 was used for color development.
Description
Background
Biotin is a water-soluble B complex vitamin (vitamin H or B7) that serves as a coenzyme in various metabolic functions. It transfers carbon dioxide for five carboxylase enzymes and is involved in the TCA cycle and gluconeogenesis. In addition, all cells require biotin for cell proliferation, production of fatty acids, and metabolism of fats and amino acids (1). Biotin recycling and attachment to histone proteins is catalyzed by an enzyme named biotinidase. Histone biotinylation is involved in regulating telomere attrition and cellular senescence. Biotinidase deficiency is a rare genetic disorder that can cause neurological and cutaneous symptoms if left untreated (2). Biotin has a high affinity for its natural ligand, avidin. The complex and irregular structure of the biotin-binding site makes it highly optimized for biotin binding and confers great specificity to the avidin-biotin complexes (3). This remarkable affinity has been exploited for numerous immunoassays including ELISA, Flow Cytometry, Immunofluorescence, In Situ Hybridization, Affinity Chromatography, and Immunohistochemistry. Anti-Biotin (D5A7) Rabbit mAb can be used as an alternative to avidins in order to minimize background and maximize signal intensity.
- Zempleni, J. et al. (2010) Biofactors 35, 36-46.
- Pindolia, K. et al. (2011) Mol Genet Metab 102, 161-9.
- Reznik, G.O. et al. (1998) Proc Natl Acad Sci USA 95, 13525-13530.
Application References
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Companion Products
For Research Use Only. Not For Use In Diagnostic Procedures.
